GAPDH, NtOSAK and CDC48, a conserved chaperone-like AAA-ATPase, as nitric oxide targets in response to (a)biotic stresses

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GAPDH, NtOSAK and CDC48, a conserved

chaperone-like AAA-ATPase, as nitric oxide targets in response to (a)biotic stresses

Jérémy Astier, Izabella Wawer, Angelique Besson-Bard, Olivier Lamotte, Sylvain Jeandroz, Herman Terenzi, Grazyna Dobrowoslka, David Wendehenne

To cite this version:

Jérémy Astier, Izabella Wawer, Angelique Besson-Bard, Olivier Lamotte, Sylvain Jeandroz, et al..

GAPDH, NtOSAK and CDC48, a conserved chaperone-like AAA-ATPase, as nitric oxide targets in response to (a)biotic stresses. 7. International Conference on the Biology, Chemistry and Therapeutic Application of Nitric Oxide, Nitric Oxide Society, Canada., Jul 2012, Édimbourg, United Kingdom.

pp.S9, �10.1016/j.niox2012.04.035�. �hal-02015252�

Thursday 26th July 2012

Session: NO in plant signaling

IS-23

S-Nitrosylation in plant defense Jörg Durner

Institute for Biochemical Plant Pathology, Helmholtz Zentrum München – German Research Center for Environmental Health, Munich-Neuherberg, Germany

Nitric oxide (NO) is an important signalling molecule in ani- mal and plant defense responses. In the latter, the redox-active molecule NO has an essential role in restriction of pathogen attack by induction of defense genes and programmed host cell death. NO can react with the thiol group of cysteine residues to form S-nitrosothiols (S-nitrosylation). A number of NO-affected proteins in plants seem to be regulated by S-nitrosylation making this type of protein modification a predominant mechanism in NO-signalling. Recently, we demonstrated that NO is a redox reg- ulator of a central transcriptional system for systemic acquired resistance in Arabidopsis thaliana. A change in the cellular redox status during the salicylic acid-mediated activation of defense leads to S-nitrosylation of the regulatory protein NPR1 and to its active monomeric form. Subsequently, the NPR1 monomers are translo- cated into the nucleus, where they interact with defense-associated transcription factors. Additional examples and evidences for NO- dependent modification of cysteine residues, describing its chemis- try/formation, specificity, and possible physiological functions in plants will be discussed. The presentation underlines the importance of NO as a redox regulator.

http://dx.doi.org/10.1016/j.niox.2012.04.033

IS-24

Nitric oxide produced during the hypersensitive response mod- ulates the plant signaling network and inhibits the pathogen’s virulence machinery

T. Ling, E. Vandelle, D. Bellin, K. Kleinfelder-Fontanesi, J.J. Huang, J.

Chen, A.M. Digby, M. Delledonne

Department of Biotechnology, University of Verona, Italy

Nitric oxide (NO) is a well-recognized key signaling mediator of the plant defense response to pathogens. Like in animals, NO acts directly or through mediators of its activity such as reactive NO derivatives (RNS) and second messengers like cGMP.

We aim at deciphering the ‘‘signaling network’’ mediated in plant by NO and RNS upon pathogen attack, and by using the HKGreen-2 dye we demonstrated that peroxynitrite, the RNS originated by the reaction between NO and O

, accumulates in plants during the hypersensitive response (HR). Peroxyni- trite is not a ‘‘death messenger’’ in plants, and its accumula- tion in plants correlates with an increase in tyrosine nitrated proteins, and we demonstrated that in vitro peroxynitrite targets specifically some MAPK kinases, inhibiting their activity and thus precisely modulating the most complex plant signal transduction network by tyrosine nitration. Furthermore, we recently found that NO can participate in plant defenses also by inhibiting pathogen effector activity. The effector HopAI1, a phosphothreonine lyase pro- duced by many Pseudomonas syringae strains which suppresses plant

immunity via MAPK inhibition, is a target of NO and its activity is dramatically inhibited by S-nitrosylation. Whereas HopAI1 expressed in Arabidopsis thaliana does not affect the HR induced by the avirulent P. syringae AvrRpt2, the expression of a mutated (Cys free) form of HopAI1 strongly reduces the hypersensitive cell death as well as plant resistance. This suggests that NO modulates the plant signaling network through nitration of specific MAPK and inhibits pathogen’s effectors that would attenuate the plant defence response.

http://dx.doi.org/10.1016/j.niox.2012.04.034

IS-25

GAPDH, NtOSAK and CDC48, a conserved chaperone-like AAA-ATPase, as nitric oxide targets in response to (a)biotic stresses

J. Astier

, I. Wawer

, A. Besson-Bard

, L. Olivier

, S. Jeandroz

, H. Terenzi

, G. Dobrowoslka

, D. Wendehenne

a

UMR 1347 Agroécologie, Pôle Mécanisme et Gestion des Interactions Plantes-microorganismes – ERL CNRS 6300, 21065 Dijon cédex, France,

b

Institute of Biochemistry and Biophysics, Polish Academy of Sciences, ul. Pawinskiego 5a, 02-106 Warsaw, Poland,

c

Centro de Biologia Molecular Estrutural Departemento de Bioquimica CCB, Univeridade Federal de Santa Catarina, 88040900 Florianopolis, SC, Brasil

Increasing evidences support the assumption that nitric oxide (NO) acts as a physiological mediator in plants facing (a)biotic stres- ses [1,2]. Understanding its effects requires a deep analysis of the molecular mechanisms underlying its mode of action. In the recent years, efforts have been made in identifying and understanding the function of plant proteins regulated by NO at the post-translational level, notably by S-nitrosylation [3].

We demonstrated that the glycolytic enzyme GAPDH undergoes a fast and transient S-nitrosylation in tobacco cells exposed to a salt stress [4]. S-nitrosylation affects only a small proportion of the GAP- DH population and does not affect glycolysis. Interestingly, in vivo GAPDH interacts with the protein kinase NtOSAK (Nicotiana tabacum osmotic stress-activated protein kinase), a member of the SnRK2 protein kinase family previously shown to be rapidly activated through NO in response to (a)biotic stresses [5]. Our current hypoth- esis is that S-nitrosylated GAPDH might acts as a phosphor-relay recruiting protein substrates for NtOSAK.

Besides GAPDH, we identified proteins undergoing S-nitrosyla- tion in tobacco cell suspensions exposed to cryptogein, a 10 kDa pro- tein produced by the oomycete Phytophthora cryptogea [6]. These proteins include CDC48, a conserved chaperone-like AAA ATPases.

Using a combination of structural and biochemical analysis, we pro- vided evidence that NO induces a local conformational change with- in the protein and inhibits its enzymatic activity. The physiological incidence of this process will be discussed.

References

[1] Besson-Bard et al.. Annu. Rev. Plant Biol. 2008;59:21–39.

[2] Rasul et al., Plant Cell Environ., in press.

[3] Astier et al.. Plant Sci. 2011;181:527–33.

[4] Wawer et al.. Biochem J. 2010;429:73–83.

[5] Lamotte et al.. Free Rad. Biol. Med. 2006;40:1369–76.

[6] Astier et al., submitted for publication.

http://dx.doi.org/10.1016/j.niox.2012.04.035

Abstracts / Nitric Oxide 27 (2012) S2–S50

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